%0 Journal Article
%T 越桔ISSR-PCR反应体系建立与优化<br>Establishment and Optimization of an ISSR-PCR Reaction System for <i>Vaccinium</i> Linn.
%A 谌月
%A 鲁艳
%A 李凌&lt
%A br&gt
%A CHEN Yue
%A LU Yan
%A LI Ling
%J 西南大学学报(自然科学版)
%D 2017
%R 10.13718/j.cnki.xdzk.2017.09.005
%X 以改良CTAB法提取越桔(<i>Vaccinium</i> Linn.)品种薄雾DNA为材料，采用正交设计L<sub>16</sub>(4<sup>5</sup>)表探讨了Tag酶；Mg<sup>2+</sup>；DNA；dntps及引物浓度5个主要因素对越桔ISSR-PCR扩增反应的影响，通过直观分析和方差分析结合，建立了越桔ISSR-PCR(20 μL)最佳反应体系，各组分的浓度为：Tag酶0.75 U，Mg<sup>2+</sup>1.875 mmol/L，DNA 0.8 mmol/L，dntp 0.1 mmol/L，引物0.2 μmol/L.在此基础上探讨了引物UBC835的最佳退火温度、循环次数和延伸时间，结果分别为54.2 ℃、35次、60 s.选用两个引物对21份越桔资源进行ISSR扩增，结果显示优化的体系稳定性较高.<br>Based on the genomic DNA extracted from <i>Vaccinium</i> Linn. cultivar Misty with a modified CTAB method, five factors influencing ISSR were optimized in an orthogonal-design experiment. Through intuitive analysis and variance analysis, an optimal ISSR-PCR reaction system was established for this fruit crop, namely 10×buffer 2 μL; 0.75 U Taq DNA polymerase; 1.875 mmol/L Mg<sup>2+</sup>; 80 ng template DNA; 0.1 mmol/L dNTPs; and 0.2 μmol/L primer. The optimal annealing temperature, number of cycles and extension time of primer UBC835 were investigated, and the results were 54.2 ℃, 35 cycles and 60s, respectively. Twenty-one <i>Vaccinium</i> accessions were used to test the stability of the optimized reaction system. The results indicated that the optimized reaction system was very stable
%K 越桔
%K 简单序列重复区间(ISSR)
%K 正交设计
%K 直观分析
%K 方差分析&lt
%K br&gt
%K &lt
%K i&gt
%K Vaccinium&lt
%K /i&gt
%K Linn
%K inter simple sequence repeat(ISSR)
%K orthogonal design
%K intuitive analysis
%K variance analysis
%U http://xbgjxt.swu.edu.cn/jsuns/html/jsuns/2017/9/201709005.htm